Identification of Scolopendra Dispensing Granules by Allele-specific PCR / 中国实验方剂学杂志

ObjectiveTo establish an allele-specific polymerase chain reaction （PCR） method for identifying Scolopendra dispensing granules， so as to ensure the quality and therapeutic effects of Scolopendra and its preparations. MethodThe primer interval suitable for the PCR was selected based on the cytochrome c oxidase subunit 3（COX-3） gene sequence of Scolopendra， and the single nucleotide polymorphism （SNP） loci of Scolopendra and its adulterants were mined from the interval for the design of specific primers. The samples of Scolopendra and its adulterants were collected. The PCR system was established and optimized regarding the annealing temperature， cycles， Taq enzymes， DNA template amount， PCR instruments， and primer concentrations， and the specificity and applicability of this method were evaluated. ResultThe PCR system was composed of 12.5 μL 2×M5 PCR Mix， 0.4 μL forward primer （10 μmol·L-1）， 0.4 μL reverse primer （10 μmol·L-1）， 2.5 μL DNA template， and 9.2 μL sterile double distilled water. PCR parameters： Pre-denaturation at 94 ℃ for 3 min， 30 cycles （94 ℃ for 20 s， 62 ℃ for 20 s， 72 ℃ for 45 s）， and extension at 72 ℃ for 5 min. After PCR amplification with the system and parameters above， the electrophoresis revealed a bright band at about 135 bp for Scolopendra and no band for the adulterants. ConclusionThe established allele-specific PCR method can accurately identify the medicinal materials， decoction pieces， and standard decoction freeze-dried powder of Scolopendra， as well as the intermediates and final products of Scolopendra dispensing granules， which is of great significance for ensuring the quality and clinical efficacy of Scolopendra and its preparations.

PCR-RFLP for Distinguishing Periplocae Cortex from Acanthopanacis Cortex and Lycii Cortex / 中国实验方剂学杂志

ObjectiveTo establish a polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） method for rapid distinguishing Periplocae Cortex from Acanthopanacis Cortex and Lycii Cortex， so as to avoid the influence of genetic confusion on drug safety. MethodThe DSS-tagged sequences of Periplocae Cortex were obtained from the Chloroplast Genome Information Resource （CGIR） and analyzed to find the enzymatic cleavage sites that were different from those of Acanthopanacis Cortex and Lycii Cortex. The specific enzymatic cleavage site， Cla I， of Periplocae Cortex was selected， on the basis of which the primers for PCR-RFLP were designed. Furthermore， the factors such as annealing temperature， number of cycles， Taq enzyme， PCR instruments， and enzymatic treatment time that may influence PCR-RFLP were studied. The established PCR-RFLP method was applied to the identification of Periplocae Cortex， Acanthopanacis Cortex， and Lycii Cortex samples produced in different regions. ResultThe PCR-RFLP at the annealing temperature of 59 ℃ and with 40 cycles showed clear bands of the samples. When the enzyme digestion time was 30 min. The reaction produced the target bands at about 140 bp and 290 bp for both Periplocae Cortex and its original plant and only a band at about 430 bp for Acanthopanacis Cortex， Lycii Cortex， and their original plants. The method can accurately distinguish Periplocae Cortex from its confounders Acanthopanacis Cortex and Lycii Cortex. ConclusionThe PCR-RFLP method for distinguishing Periplocae Cortex from Acanthopanacis Cortex and Lycii Cortex was established. It has high stability， sensitivity， and applicability， providing a reference for the quality control of Periplocae Cortex， Acanthopanacis Cortex， and Lycii Cortex.

Identification of Original Plants of Lycii Cortex by Multiplex Allele-specific PCR / 中国实验方剂学杂志

ObjectiveTo identify Lycium chinense and L. barbarum as the original plants of Lycii Cortex simply and efficiently by multiple allele-specific polymerase chain reaction （PCR）. MethodThe chloroplast genome sequences of L. chinense and L. barbarum were downloaded from the Chloroplast Genome Information Resource （CGIR）， and then IdenDSS was employed to screen out the specific single nucleotide polymorphism （SNP） sites between the two plants. Primer 5.0 was used to design the specific primers， including primers GQ-F/R for identifying L. chinense and primers NX-F/R for identifying L. barbarum. Furthermore， the primer concentration ratio， annealing temperature， cycles， and Taq enzyme were optimized to establish the optimal PCR system and conditions for plant identification. Finally， the applicability of the established method was examined with the plant samples collected from different regions. ResultThe PCR with GQ-F/R∶NX-F/R concentration ratio of 2∶1 at the annealing temperature at 59 ℃ and for 30 cycles showed specific bands at 183 bp and 295 bp， respectively， for L. chinense and L. barbarum samples from different regions. ConclusionThe established PCR approach can simply， rapidly， and efficiently identify the original plants of Lycii Cortex， serving as a new method for the discrimination between L. chinense and L. barbarum. Moreover， the method provides technical support for the research and development of classic famous prescriptions containing Lycii Cortex.

Specific PCR for Identification of Astragalus membranaceus var. mongholicus Seeds， A. membranaceus Seeds， and Adulterants / 中国实验方剂学杂志

ObjectiveTo establish a method based on specific polymerase chain reaction （PCR） that can accurately and rapidly identify Astragalus membranaceus var. mongholicus （AMM） seeds and A. membranaceus （AM） seeds. MethodThe Chloroplast Genome Information Resource （CGIR） and IdenDSS were used to obtain the characteristic DNA fragments of AMM and AM， and the specific single nucleotide polymorphism （SNP） sites of AMM and AM were screened out， on the basis of which the specific primers MG-F/MG-R of AMM and MJ-F/MJ-R of AM were designed. The specific PCR method for identifying AMM and AM was established and optimized， and the specificity and applicability of the method were investigated. ResultThe specific PCR conditions for the identification of AMM were primers MG-F/MG-R， annealing at 62 ℃， and 28 cycles. After PCR amplification and gel electrophoresis， the specific band appeared at about 220 bp， with no band for the seeds of AM or adulterants. The specific PCR conditions for identifying the AM were primers MJ-F/MJ-R， annealing at 58 ℃， and 28 cycles. After PCR amplification and gel electrophoresis， the band appeared at about 150 bp， with no band of AMM or adulterants. ConclusionThe specific PCR method established in this study can accurately and quickly identify the seeds of AMM and AM， which provides a basis for the classification and accurate identification of Astragalus seeds and adulterants.

ABC-AF-Stroke score predicts thromboembolism in non-anticoagulated patients following successful atrial fibrillation ablation: a report from the Chinese Atrial Fibrillation Registry / 中华医学杂志(英文版)

BACKGROUND@#The age, biomarkers, and clinical history (ABC)-atrial fibrillation (AF)-Stroke score have been proposed to refine stroke risk stratification, beyond what clinical risk scores such as the CHA2DS2-VASc score can offer. This study aimed to identify risk factors associated with thromboembolism and evaluate the performance of the ABC-AF-Stroke score in predicting thromboembolism in non-anticoagulated AF patients following successful ablations.@*METHODS@#A total of 2692 patients who underwent successful ablations with discontinued anticoagulation after a 3-month blanking period in the Chinese Atrial Fibrillation Registry (CAFR) between 2013 and 2019 were included. Cox regression analysis was conducted to present the association of risk factors with thromboembolism risk. The ABC-AF-Stroke score was evaluated in terms of discrimination, including concordance index (C-index), net reclassification improvement (NRI) and integrated discrimination improvement (IDI), clinical utilization by decision curve analysis (DCA), and calibration by comparing the predicted risk with the observed annualized event rate.@*RESULTS@#After a median follow-up of 3.5 years, 64 patients experienced thromboembolism events. Age, prior history of stroke/transient ischemic attack (TIA), high-sensitivity cardiac troponin T (cTnT-hs), and N-terminal pro-B-type natriuretic peptide (NT-proBNP) were independently associated with thromboembolism risk. The ABC-AF-Stroke score performed statistically significantly better than the CHA2DS2-VASc score in terms of C-index (0.67, 95% confidence interval [CI]: 0.59-0.74 vs. 0.60, 95% CI: 0.52-0.67, P = 0.030) and reclassification capacity. The DCA implied that the ABC-AF-Stroke score could identify more thromboembolism events without increasing the false positive rate compared to the CHA2DS2-VASc score. The calibration curve showed that the ABC-AF-Stroke score was well calibrated in this population.@*CONCLUSIONS@#In this real-world study enrolling non-anticoagulated AF patients following successful ablations, age, prior history of stroke/TIA, level of NT-proBNP, and cTnT-hs were independently associated with an increased risk of thromboembolism. The ABC-AF-Stroke score was well-calibrated and statistically significantly outperformed the CHA2DS2-VASc score in predicting thromboembolism risk.

Leukemia Genotype Analysis of Children with Acute Lymphoblastic Leukemia in Yunnan Area / 中国实验血液学杂志

OBJECTIVE@#To analyze 43 leukemia genes in children with acute lymphoblastic leukemia (ALL) in Yunnan province, and provide the basis for the diagnosis and treatment of children with ALL in this area.@*METHODS@#The clinical data of 428 children with newly diagnosed ALL in Yunnan area from January 2015 to December 2020 were retrospectively analyzed. Multiple nested PCR technology was used to detect 43 common leukemia genes.@*RESULTS@#Among the 428 children with ALL, 159 were positive for leukemia genes, with a positive rate of 37.15% (159/428), and a total of 15 leukemia genes were detected. Among the 159 leukemia gene-positive children, ETV6-RUNX1+ accounted for 25.79% (41/159), followed by E2A-PBX1+ and BCR-ABL+, accounting for 24.53% (39/159) and 23.27% (37/159) respectively. MLL+ accounted for 6.29% (10/159), WT1+ accounted for 4.40% (7/159), IKZF1 gene deletion and CRLF2+ accounted for 3.77% (6/159) respectively. The positive rate of MLL (46.15%) was the highest in ＜1-year old group, the positive rate of ETV6-RUNX1 (10.56%) was the highest in 1-10-year old group, and BCR-ABL+ rate (23.65%) was the highest in >10-year old group. The distribution of leukemia genes in different age groups was statistically significant (P <0.05).@*CONCLUSION@#The most common fusion gene of children with ALL in Yunnan is ETV6-RUNX1, followed by E2A-PBX1 and BCR-ABL.

Imputation method for dropout in single-cell transcriptome data / 生物医学工程学杂志

Single-cell transcriptome sequencing (scRNA-seq) can resolve the expression characteristics of cells in tissues with single-cell precision, enabling researchers to quantify cellular heterogeneity within populations with higher resolution, revealing potentially heterogeneous cell populations and the dynamics of complex tissues. However, the presence of a large number of technical zeros in scRNA-seq data will have an impact on downstream analysis of cell clustering, differential genes, cell annotation, and pseudotime, hindering the discovery of meaningful biological signals. The main idea to solve this problem is to make use of the potential correlation between cells and genes, and to impute the technical zeros through the observed data. Based on this, this paper reviewed the basic methods of imputing technical zeros in the scRNA-seq data and discussed the advantages and disadvantages of the existing methods. Finally, recommendations and perspectives on the use and development of the method were provided.

A novel biological sources consistency evaluation method reveals high level of biodiversity within wild natural medicine: A case study of Amynthas earthworms as "Guang Dilong"

For wild natural medicine, unanticipated biodiversity as species or varieties with similar morphological characteristics and sympatric distribution may co-exist in a single batch of medical materials, which affects the efficacy and safety of clinical medication. DNA barcoding as an effective species identification tool is limited by its low sample throughput nature. In this study, combining DNA mini-barcode, DNA metabarcoding and species delimitation method, a novel biological sources consistency evaluation strategy was proposed, and high level of interspecific and intraspecific variations were observed and validated among 5376 Amynthas samples from 19 sampling points regarded as "Guang Dilong" and 25 batches of proprietary Chinese medicines. Besides Amynthas aspergillum as the authentic source, 8 other Molecular Operational Taxonomic Units (MOTUs) were elucidated. Significantly, even the subgroups within A. aspergillum revealed here differ significantly on chemical compositions and biological activity. Fortunately, this biodiversity could be controlled when the collection was limited to designated areas, as proved by 2796 "decoction pieces" samples. This batch biological identification method should be introduced as a novel concept regarding natural medicine quality control, and to offer guidelines for in-situ conservation and breeding bases construction of wild natural medicine.

Identification of Arisaematis Rhizoma and Pinelliae Rhizoma by PCR-RFLP / 中国实验方剂学杂志

ObjectiveIn recent years， with the sharp decline of wild resources in Arisaematis Rhizoma and Pinelliae Rhizoma and the immaturity of medicinal cultivation technology， their adulterants have appeared frequently in the market， and the main identifying characteristics have mostly disappeared in the circulation of medicinal materials. Therefore， there is an urgent need to establish a molecular identification method that can quickly and effectively identify the specificity of Arisaematis Rhizoma and Pinelliae Rhizoma. MethodAfter comparison of the rbcL sequences of Arisaematis Rhizoma，Pinelliae Rhizoma， and their adulterants， the specific enzyme cleavage sites Hae Ⅲ and Dra Ⅰ of Arisaematis Rhizoma and Pinelliae Rhizoma， respectively， were selected and identified by polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP）. The main system conditions of PCR-RFLP reaction were established and optimized， and their durability and the ability to detect genuine， adulterants， and mixed counterfeits were investigated. ResultThe PCR-RFLP identification method of Arisaematis Rhizoma and Pinelliae Rhizoma was established. After specific primer amplification， Arisaematis Rhizoma and Pinelliae Rhizoma could be digested by Hae Ⅲ and Dra Ⅰ-restricted endonucleases respectively， at annealing temperature of 54 ℃， the number of cycles of 35， and the amount of DNA template of 3-30 ng， producing two fragments or small cut fragments with a single band between 100-250 bp， whereas the mixed counterfeits were not cleaved and both showed a band at 250 bp. The method is highly accurate in identifying adulterants and mixed counterfeits of Arisaematis Rhizoma or Pinelliae Rhizoma. ConclusionThe PCR-RFLP method developed in this study allows for the rapid identification of Arisaematis Rhizoma and Pinelliae Rhizoma.

Catheter ablation versus medical therapy for atrial fibrillation with prior stroke history: a prospective propensity score-matched cohort study / 老年心脏病学杂志（英文版）

BACKGROUND@#Patients with atrial fibrillation (AF) and prior stroke history have a high risk of cardiovascular events despite anticoagulation therapy. It is unclear whether catheter ablation (CA) has further benefits in these patients.@*METHODS@#AF patients with a previous history of stroke or systemic embolism (SE) from the prospective Chinese Atrial Fibrillation Registry study between August 2011 and December 2020 were included in the analysis. Patients were matched in a 1:1 ratio to CA or medical treatment (MT) based on propensity score. The primary outcome was a composite of all-cause death or ischemic stroke (IS)/SE.@*RESULTS@#During a total of 4.1 ± 2.3 years of follow-up, the primary outcome occurred in 111 patients in the CA group (3.3 per 100 person-years) and in 229 patients in the MT group (5.7 per 100 person-years). The CA group had a lower risk of the primary outcome compared to the MT group [hazard ratio (HR) = 0.59, 95% CI: 0.47-0.74, P < 0.001]. There was a significant decreasing risk of all-cause mortality (HR = 0.43, 95% CI: 0.31-0.61, P < 0.001), IS/SE (HR = 0.73, 95% CI: 0.54-0.97, P = 0.033), cardiovascular mortality (HR = 0.32, 95% CI: 0.19-0.54, P < 0.001) and AF recurrence (HR = 0.33, 95% CI: 0.30-0.37, P < 0.001) in the CA group compared to that in the MT group. Sensitivity analysis generated consistent results when adjusting for time-dependent usage of anticoagulants.@*CONCLUSIONS@#In AF patients with a prior stroke history, CA was associated with a lower combined risk of all-cause death or IS/SE. Further clinical trials are warranted to confirm the benefits of CA in these patients.

Correlation between cognitive impairment and cortical atrophy in elderly patients with asymptomatic carotid artery stenosis / 中华健康管理学杂志

Objective:To analyze the correlation between cognitive impairment and cortical atrophy in elderly patients with asymptomatic carotid artery stenosis (ACAS).Methods:In this cross-sectional study, 40 consecutive elderly patients with ACAS treated in the Department of Neurology, Northern Jiangsu People′s Hospital from July 1, 2020 to June 30, 2021 (ACAS group), and 40 elderly healthy controls who accepted physical examination during the same period (control group) were included. Cognitive assessment was performed using the Mental State Examination Scale (MMSE) and the Montreal Cognitive Assessment Scale (MoCA), and brain magnetic resonance imaging scanning was performed in the ACAS group. The artificial intelligence technique was applied for brain lobe segmentation and cortical volume calculation. The χ2-test, independent sample t-test and Wilcoxon non-parametric test were used to analyze the difference of clinical data and cognitive scores between the two groups. In the ACAS group, the cortical volumes of the side with carotid stenosis was compared with that of the normal side, and Spearman′s correlation analysis was used to assess the correlation between cognitive scores and cortical atrophy. Results:Compared with the control group, the ACAS group got significantly lower scores of MMSE and MoCA, as well as lower scores of visuospatial executive function, attention and calculation, language function, abstraction ability and delayed recall [(25.60±2.49) vs (27.18±1.01), (22.05±3.59) vs (25.60±1.43), (2.73±1.04) vs (4.08±0.62), (4.53±0.93) vs (5.03±0.66), 2.00 (0.00) vs 3.00 (0.00), 1.00 (1.00) vs 2.00 (0.00), and (2.95±0.96) vs (3.35±0.62)] (all P<0.05). There was not significant differences in naming and orientation ability between the two groups (both P>0.05). The volume of cortical, temporal lobe, frontal lobe, parietal lobe and insular lobe on the side with carotid stenosis in the ACAS group were significantly smaller than those on the normal side [186.23 (177.97, 202.53) vs 194.67 (185.65, 204.82) cm 3, 54.74 (50.66, 56.95) vs 55.61 (51.24, 58.49) cm 3, 72.98 (70.76, 78.34) vs 75.27 (72.34, 80.66) cm 3, 53.66 (51.11, 57.86) vs 56.59 (52.80, 60.09) cm 3, 6.57 (6.35, 7.07) vs 6.72 (6.46, 7.34) cm 3] (all P<0.05). The MoCA score in the ACAS group was positively related to the cortical volume ratio of the two sides ( r=0.427, P<0.01). The attention ( r=0.353) and abstraction ( r=0.226) ability scores were positively correlated with the temporal lobe volume ratios of the two sides (both P<0.05). The visuospatial executive ( r=0.187) and language ( r=0.373) ability scores were positively correlated with frontal lobe volume ratios of the two sides (both P<0.05), and visuospatial executive ( r=0.386), naming ( r=0.344), language ( r=0.517), abstraction ( r=0.335) and delayed recall ( r=0.333) ability scores were positively correlated with parietal lobe volume ratios of the two sides (all P<0.05). Conclusion:In elderly patients with ACAS, the cognitive impairment and cortical atrophy on the sides with carotid stenosis are significant and a positive correlation is detected between them.

Clinical effects of treatment of single-segment lumbar tuberculosis by oblique lateral interbody fusion and percutaneous pedicle screw fixation / 西安交通大学学报(医学版)

【Objective】 To investigate the clinical effects of treatment of single-segment lumbar tuberculosis by oblique lateral interbody fusion with autologous iliac bone and percutaneous pedicle screw fixation. 【Methods】 We collected the clinical data of 47 patients with lumbar tuberculosis treated in The First Affiliated Hospital of Xi’an Jiaotong University from March 2017 to January 2020. Among them， 22 patients underwent oblique lateral interbody fusion with autologous iliac bone and percutaneous pedicle screw fixation （minimally invasive group） and 25 patients underwent open surgery combined anterior-debridement and posterior-fixation （control group）. The related data were collected， including gender， sex， body mass index （BMI）， systemic symptoms of tuberculosis， operation duration， intraoperative bleeding， postoperative drainage， hospital stay， complications， visual analogue score （VAS）， erythrocyte sedimentation rate （ESR）， and Oswestry disability index （ODI）. 【Results】 Baseline clinical characteristics did not significantly differ between the two groups （P>0.05）. Compared with control group， the minimally invasive group had shorter operation duration ［（188.64±18.59） min vs. （201.60±22.67） min］， less intraoperative blood loss ［（118.64±22.95） mL vs. （553.60±100.54） mL］， less postoperative drainage ［（134.55±36.48） mL vs. （291.20±61.53） mL］， and shorter hospitalization time ［（12.86±2.17） d vs. （15.80±3.03） d］ （all P0.05）. Compared with the preoperative ones， ESR， VAS score and ODI score significantly decreased and Cobb angle significantly increased in both groups （all P0.05）. 【Conclusion】 Both minimally invasive technique and open surgery can achieve excellent clinical results， but the minimally invasive technique can reduce the surgical trauma and shorten the hospitalization time.

OShnscc: a novel user-friendly online survival analysis tool for head and neck squamous cell carcinoma based on RNA expression profiles and long-term survival information / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Head and neck squamous cell carcinoma (HNSCC), as the most common type (>90%) of head and neck cancer, includes various epithelial malignancies that arise in the nasal cavity, oral cavity, pharynx, and larynx. In 2020, approximately 878 ‍ 000 new cases and 444 000 deaths linked to HNSCC occurred worldwide (Sung et al., 2021). Due to the associated frequent recurrence and metastasis, HNSCC patients have poor prognosis with a five-year survival rate of 40%-50% (Jou and Hess, 2017). Therefore, novel prognostic biomarkers need to be developed to identify high-risk HNSCC patients and improve their disease outcomes.

Clinical effects of robot-assisted minimal invasive transforaminal lumbar interbody fusion in the treatment of single-segment lumbar disc herniation / 西安交通大学学报(医学版)

【Objective】 To compare the clinical effects and screw placement accuracy for treating lumbar disc herniation between robot-assisted minimal invasive transforaminal lumbar interbody fusion (RA-MIS-TLIF) and minimal invasive transforaminal lumbar interbody fusion (MIS-TLIF). 【Methods】 We retrospectively recruited 69 patients with single segment lumbar disc herniation treated between January 2018 and August 2019 at Honghui Hospital of Xi’an Jiaotong University. There were cases of 33 RA-MIS-TLIF (RA group) and 36 MIS-TLIF (MIS-TLIF group). Subsequently, the patients’ baseline characteristics were collected, including age, gender, body mass index, complication with diabetes, duration of symptoms, operated segment, and follow-up time. We also collected perioperative parameters such as operation time, intraoperative blood loss, intraoperative fluoroscopy frequency, screw placement accuracy, wound drainage, hospitalization duration, postoperative complicatins, and fusion rate. Lower back pain, lower extremity pain visual analogue score (VAS), and lumbar Japanese Orthopaedic Association Scores (JOA) were obtained preoperatively, postoperative 3 days/6 months/12 months, and the last follow-up. 【Results】 All the procedures were successfully completed and the follow-up time was 14.82±1.83 (RA group) and 15.11±1.62 (MIS-TLIF group) months, without significant difference (P>0.05). Compared with MIS-TLIF group, RA group had less intraoperative blood loss [(116.67±18.48) min vs. (128.06±22.53) min], fluoroscopy frequency [(12.42±2.28) vs. (15.67±2.46)], screw placement accuracy (93.18% vs. 84.03%), postoperative drainage [(73.03±23.52) mL vs. (88.33±28.54) mL], and shorter hospitalization stay [(6.45±1.52)d vs. (7.69±1.85) d] (all P0.05). The VAS of lower back pain and lower extremity pain, and lumbar JOA were significantly improved after the operation (P0.05). Meanwhile, fusion rate and incidence of complications did not significantly differ between the two groups (P>0.05). 【Conclusion】 Both robot-assisted MIS-TLIF and MIS-TLIF can achieve excellent clinical effects in treating single-segment lumbar disc herniation. However, the former can improve the accuracy of screw placement and reduce intraoperative blood loss, fluoroscopy frequency, postoperative drainage and hospitalization time, which indicates a promising application.

Conversion therapy strategies for hepatocellular carcinoma / 中华消化外科杂志

Hepatectomy is the main optimal curative treatment of hepatocellular carci-noma (HCC) to achieve long-term survival. However, most patients in China do not fulfill the criteria for surgery due to the intermediate-advanced stage of HCC at their initial diagnosis. With the promising advances in locoregional and systematic therapies, development of targeted drugs, success of immunotherapy, as well as the emergence of the therapeutic alliance, conversion therapy has well developed nowadays and become a hotspot in recent years. A part of unresectable HCC patients have afforded sequent radical surgery opportunities and prolonged the overall survival through improving liver function, increasing the residual liver volume, and minimizing tumor volume. At present, target therapy combined immunotherapy, local therapy combined systemic therapy are commonly used and widely applicable conversion therapy modes in China. Based on expansion of conversion therapy concepts, more high-level evidences are needed to exploit the full potential of conversion treatment strategies, accurately select candidates, determine the timing of surgery, improve conversion rate, guarautee the safety and long-term efficacy, which requires further investigation and research.

Clinical trials on intrathecal pemetrexed treated leptomeningeal metastases from solid tumors / 中华肿瘤杂志

Objective: To investigate the feasibility, safety and efficacy of intrathecal pemetrexed (IP) treated for patients with leptomeningeal metastases (LM) from solid tumors. Methods: Forty-seven patients receiving pemetrexed intrathecal chemotherapy in the First Hospital of Jilin University from 2017 to 2018 were selected. The study of pemetrexed intrathecal chemotherapy adopted the classical dose-climbing model and included 13 patients with meningeal metastasis of non-small cell lung cancer who had relapsed and refractory after multiple previous treatments including intrathecal chemotherapy. Based on the dose climbing study, 34 patients with meningeal metastasis of solid tumor who did not receive intrathecal chemotherapy were enrolled in a clinical study using pemetrexed as the first-line intrathecal chemotherapy combined with radiotherapy. Kaplan-Meier method and Log rank test were used for survival analysis, and Cox regression model was used for influencing factor analysis. Results: The dose climbing study showed that the maximum tolerated dose of pemetrexed intrathecal chemotherapy was 10 mg per single dose, and the recommended dosing regimen was 10 mg once or twice a week. The incidence of adverse reactions was 10 cases, including hematological adverse reactions (7 cases), transaminase elevation (2 cases), nerve root reactions (5 cases), fatigue and weight loss (1 case). The incidence of serious adverse reactions was 4, including grade 4-5 poor hematology (2 cases), grade 4 nerve root irritation (2 cases), and grade 4 elevated aminotransferase (1 case). In the dose climbing study, 4 patients were effectively treated and 7 were disease controlled. The survival time was ranged from 0.3 to 14.0 months and a median survival time was 3.8 months. The clinical study of pemetrexed intrathecal chemotherapy combined with radiotherapy showed that the treatment mode of 10 mg pemetrexed intrathecal chemotherapy once a week combined with synchronous involved area radiotherapy 40 Gy/4 weeks had a high safety and reactivity. The incidence of major adverse reactions was 52.9% (18/34), including hematologic adverse reactions (13 cases), transaminase elevation (10 cases), and nerve root reactions (4 cases). In study 2, the response rate was 67.6% (23/34), the disease control rate was 73.5% (25/34), the overall survival time was ranged from 0.3 to 16.6 months, the median survival time was 5.5 months, and the 1-year survival rate was 21.6%. Clinical response, improvement of neurological dysfunction, completion of concurrent therapy and subsequent systemic therapy were associated with the overall survival (all P<0.05). Conclusions: Pemetrexed is suitable for the intrathecal chemotherapy with a high safety and efficacy. The recommended administration regimen was IP at 10 mg on the schedule of once or twice per week. Hematological toxicity is the main factor affecting the implementation of IP. Vitamin supplement can effectively control the occurrence of hematological toxicity.

The relationship between fasting blood glucose level and thromboembolism events in patients with non-valvular atrial fibrillation / 中华心血管病杂志

Objective: To explore the relationship between fasting blood glucose level and thromboembolism events in patients with non-valvular atrial fibrillation (NVAF). Methods: This was an observational study based on data from a multicenter, prospective Chinese atrial fibrillation registry cohort, which included 18 703 consecutive patients with atrial fibrillation (AF) in 31 hospitals in Beijing from August 2011 to December 2018. Patients were divided into 5 groups according to status of comorbid diabetes and fasting glucose levels at admission: normal blood glucose (normal glucose group), pre-diabetes group, strict glycemic control group, average glycemic control group and poor glycemic control group. Patients were followed up by telephone or outpatient service every 6 months. The primary follow-up endpoint was thromboembolic events, including ischemic stroke and systemic embolism. The secondary endpoint was the composite endpoint of cardiovascular death and thromboembolic events. Kaplan-Meier survival analysis and multifactorial Cox regression were used to analyze the correlation between fasting glucose levels and endpoint events. Results: The age of 18 703 patients with NVAF was (63.8±12.0) years, and there were 11 503 (61.5%) male patients. There were 11 877 patients (63.5%) in normal blood glucose group, 2 023 patients (10.8%)in pre-diabetes group, 1 131 patients (6.0%) in strict glycemic control group, 811 patients in average glycemic control group and 2 861 patients(4.3%) in poor glycemic control group. Of the 4 803 diabetic patients, 1 131 patients (23.5%) achieved strict glycemic control, of whom 328 (29.0%) were hypoglycemic (fasting blood glucose level<4.4 mmol/L at admission). During a mean follow-up of (51±23) months (up to 82 months), thromboembolic events were reported in 984 patients (5.3%). The survival curve analysis of Kaplan Meier showed that the incidence rates of thromboembolic events in normal glucose group, pre-diabetes group, strict glycemic control group, average glycemic control group and poor glycemic control group were 1.10/100, 1.41/100, 2.09/100, 1.46/100 and 1.71/100 person-years, respectively (χ²=53.0, log-rank P<0.001). The incidence rates of composite endpoint events were 1.86/100, 2.17/100, 4.08/100, 2.58/100, 3.16/100 person-years (χ²=72.3, log-rank P<0.001). The incidence of thromboembolic events and composite endpoint events in the other four groups were higher than that in the normal blood glucose group (P<0.001). Multivariate Cox regression analysis showed that compared with normal glucose group, the risk of thromboembolism increased in pre-diabetes group(HR=1.23, 95%CI 1.00-1.51, P=0.049), strict glycemic control group(HR=1.32, 95%CI 1.06-1.65, P=0.013) and poor glycemic control group(HR=1.26, 95%CI 1.01-1.58, P=0.044). Conclusion: Both high or low fasting glucose may be an independent risk factor for thromboembolic events in patients with NVAF.

Specific PCR Identification of Hibisci Cortex and Its Adulterants Based on DNA Signature Sequence Tags / 中国实验方剂学杂志

ObjectiveUncommon medicinal herbs are valuable medicinal resources， but their identification is a difficult problem in Chinese medicine due to their particularity and complexity. It is， therefore， urgent to establish a method for the identification of uncommon medicinal herbs. In this study， DNA signature sequence （DSS） tags were used to establish a specific polymerase chain reaction （PCR） identification method for Hibisci Cortex， the origin plant of Hibisci Cortex， and its adulterants. MethodThe candidate DSS tags were obtained from the chloroplast genome sequence analysis， and the DSS tags were verified by DNA sequencing. The specific identification primers for H. syriacus were designed based on the obtained reliable DSS tags. The PCR reaction conditions were optimized， and the tolerance and feasibility were investigated. ResultA DSS tag for identification of H. syriacus was obtained from the comparison of sequencing results of the amplified products with DSS， which revealed the distinguishing characteristics of Hibisci Cortex and its adulterants. A pair of specific primers for H. syriacus was designed according to the DSS tag. After PCR amplification and gel electrophoresis with the primers， a single bright band of about 270 bp was observed from H. syriacus， which did not appear in the four adulterants. ConclusionA DSS tag obtained in this study can be used to identify H. syriacus. The specific primers designed based on this DSS tag can accurately and simply identify the original plant of Hibisci Cortex and its adulterants， which provides a new method and idea for the molecular identification of genuine and counterfeit products of Hibisci Cortex.

Specific PCR Identification of Artemisia absinthium， A New Foreign Medicinal Resource of Artemisia / 中国实验方剂学杂志

ObjectiveTo establish a specific polymerase chain reaction （PCR） method for the identification of Artemisia absinthium to allow accurate and convenient identification of A. absinthium and its related species. MethodThe chloroplast genome sequences of A. absinthium and its related species were searched from Chloroplast Genome Information Resource （CGIR）， and the specific single nucleotide polymorphism （SNP） sites of A. absinthium were screened out. A pair of specific identification primers （zykh1-F and zykh1-R） of A. absinthium was designed. The original plant samples of A. absinthium and its related species were collected. The specific PCR method was established and optimized， and the tolerance and feasibility of this method were investigated and verified. The method was used to identify A. absinthium samples purchased from Xinjiang medicinal materials market. ResultA 210 bp bright band was obtained from A. absinthium after PCR amplification and gel electrophoresis under the following conditions： specific primers zykh1-F and zykh1-R， annealing temperature of 54 ℃， and the number of cycles of 33. No such band was observed from its relative species， such as A. argyi， A. annua， A. leucophylla， and A. lavandulaefolia. ConclusionThe specific PCR identification method of established in this study can accurately identify A. absinthium and its common related species with high specificity. The method can save time and cost and allows a convenient and fast species identification for the introduction and utilization of A. absinthium resources.

Identification of Pheretima （Pheretima aspergillum） Formula Granule by Allele-specific PCR / 中国实验方剂学杂志

ObjectiveIn order to ensure the safety and effectiveness of clinical drug use ， the identification method of mixed and adulterated specific polymerase chain reaction （PCR） identification of Pheretima aspergillum and its processed products was established. MethodBased on the cytochrome C oxidase subunit I sequence of P. aspergillum， primers were designed to cover the whole sequences， and the stable DNA ranges suitable for the identification of Pheretima （P. aspergillum） formula granule were screened out. Specific primers were designed according to the specific single nucleotide polymorphisms （SNP） of P. aspergillum in the stable DNA range. The P. aspergillum and its mixture were collected respectively， the PCR reaction system was established and optimized， and PCR reaction system and procedure were optimized， and the tolerance and applicability were investigated. ResultWhen the annealing temperature was 62 ℃ and the cycle number was 36， both P. aspergillum formula granule and its formula particles could amplify a single specific identification band of about 170 bp， and the other 20 adulterants and negative controls had no band. ConclusionThe allele-specific PCR identification method established in this study can quickly and accurately identify the P. aspergillum formula granule. The orgin of Chinese herbal medicine and decoction pieces and P. aspergillum were accurately identified. It can also provide a reference for other studies on the quality standard research of other Chinese herbal formula granule.